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bioRxiv · 10.1101/2025.10.28.685246

DNA Framework Nanoreactor for Programmable Membrane Fusion

Shi, Q.·Yang, Q.·Li, F.·Bao, M.·Wang, S.·Huang, K.·Liu, J.·Wang, Y.·Chen, Y.·Yang, Y. R.·Bian, X.·Wu, Z.·Yang, Y.
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Abstract

Membrane fusion is a fundamental yet transient process that has long resisted direct structural and kinetic dissection. Here we introduce a DNA framework vesicle (DFV) nanoreactor that transforms this elusive biological phenomenon into a programmable, visualizable, and quantifiable process-a nanoreaction confined in space yet extended in time. By embedding lipid membranes and SNARE proteins within precisely defined DNA apertures, DFVs convert stochastic vesicle collisions into geometry-controlled fusion events with tunable kinetics. Cryo-electron microscopy resolves a complete sequence of six intermediates, revealing how nanoscale confinement reshapes the energetic landscape of bilayer merger. Quantitative fluorescence and nano-flow cytometry further establish a direct link between spatial design and fusion probability. Extending this concept to living cells, DFVs enable controllable membrane fusion and augmentation on VAMP2-expressing membranes, achieving direct cytosolic delivery of functional siRNA via fusion-driven transfer rather than endocytosis. This framework bridges structural precision and functional mimicry, offering a unified platform to reconstruct, quantify, and harness membrane fusion as a programmable process for synthetic biology and nanomedicine.

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Shi, Q., Yang, Q., Li, F., Bao, M., Wang, S., Huang, K., Liu, J., Wang, Y., Chen, Y., Yang, Y. R., Bian, X., Wu, Z., Yang, Y.. 2025-10-30. DNA Framework Nanoreactor for Programmable Membrane Fusion. https://doi.org/10.1101/2025.10.28.685246

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